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A Chromosomally Based Tod-luxCDABEWhole-Cell Reporter For Benzene, Toluene, Ethybenzene, And Xylene (BTEX) Sensing

B. M. Applegate, S. R. Kehrmeyer, G. S. Sayler

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ABSTRACTAtod-luxCDABEfusion was constructed and introduced into the chromosome ofPseudomonas putidaF1, yielding the strain TVA8. This strain was used to examine the induction of thetodoperon when exposed to benzene, toluene, ethylbenzene, and xylene (BTEX) compounds and aqueous solutions of JP-4 jet fuel constituents. Since this system contained the completeluxcassette (luxCDABE), bacterial bioluminescence in response to putative chemical inducers of thetodoperon was measured on-line in whole cells without added aldehyde substrate. There was an increasing response to toluene concentrations from 30 μg/liter to 50 mg/liter, which began to saturate at higher concentrations. The detection limit was 30 μg/liter. There was a significant light response to benzene,m- andp-xylenes, phenol, and water-soluble JP-4 jet fuel components, but there was no bioluminescence response upon exposure too-xylene. The transposon insertion was stable and had no negative effect on cell growth.